BACKGROUND & AIMS: OBJECTIVE:To determine the degree of correlation between Farr and ELISA methods of detecting anti-dsDNA antibodies in patients with systemic lupus erythematosus (SLE), and their association with measures of disease activity. METHODS:Anti-dsDNA antibodies were assayed using the Farr and ELISA methods in patients followed between January 1, 2000, and December 31, 2002. Statistical correlations between Farr and ELISA were determined. Relationships between the 2 assays and measures of disease activity [SLE Disease Activity Index 2000 (SLEDAI-2K-DNA), renal, central nervous system (CNS), and vasculitis] were determined for the same clinic visit. RESULTS:550 patients with 2940 clinic visits met the inclusion criteria. Correlation between Farr and ELISA levels was 0.46 using the first visit for each patient. When the Farr was abnormal, the ELISA was equally likely to be normal or abnormal. Abnormal Farr results were associated with higher SLEDAI-2K scores than normal Farr results (6.2 vs 4.3, respectively; p < 0.0001). There was less of a distinction with ELISA results (5.9 vs 4.8; p = 0.04). Farr levels were significantly associated with the presence of renal disease and vasculitis, while ELISA levels were not. Neither Farr nor ELISA results correlated with the presence of active CNS involvement. CONCLUSION:Farr and ELISA techniques for the detection of anti-dsDNA antibodies in patients with SLE are poorly correlated. The Farr is superior to the ELISA in correlating with measures of global disease activity, as well as renal and vasculitis involvement. The Farr technique should continue to be used in clinical practice. The ELISA adds no additional information.

背景与目标：

BACKGROUND & AIMS: Although asthmatic patients are known to have increased levels of IgG antibody against house dust mite (HDM), it is not clear whether or not the presence of HDM-specific IgG antibody is associated with the etiological mechanism of asthma. To address this problem, we evaluated the relationship between HDM-specific IgG antibody levels and incidence of asthma in a general pediatric population. IgE and IgG antibody levels against Dermatophagoides farinae (Df) were examined by RAST and ELISA in a total of 722 randomly selected schoolchildren including 26 subjects with asthma, and the relative prevalence rates of asthma in this population were evaluated in relation to both Df-specific IgE and IgG levels. The incidence of asthma correlated not only with levels of Df-specific IgE, but also with those of Df-specific IgG. There was a significant correlation between Df-specific IgE and IgG levels both in the total population and in the asthmatic children. Because IgG and IgE responses occurred in parallel in this population, the clinical significance of HDM-specific IgG anti-body remains unclear. However, our findings have suggested that clinical expression of asthma in children is primarily dependent on their capacity to mount a immune response to HDM, which includes both IgE and IgG responses.

背景与目标： 尽管已知哮喘患者针对屋尘螨 (HDM) 的IgG抗体水平升高，但尚不清楚HDM特异性IgG抗体的存在是否与哮喘的病因机制有关。为了解决这个问题，我们评估了普通儿童人群中HDM特异性IgG抗体水平与哮喘发生率之间的关系。通过RAST和ELISA检测了总共722名随机选择的学童 (包括26名哮喘受试者) 的针对粉尘螨 (Df) 的IgE和IgG抗体水平，并评估了该人群中哮喘的相对患病率与Df特异性IgE和IgG水平。哮喘的发病率不仅与Df特异性IgE的水平相关，而且与Df特异性IgG的水平相关。在总人口和哮喘儿童中，Df特异性IgE和IgG水平之间存在显着相关性。由于IgG和IgE反应在该人群中并行发生，因此HDM特异性IgG抗体的临床意义尚不清楚。但是，我们的发现表明，儿童哮喘的临床表达主要取决于他们对HDM产生免疫反应的能力，其中包括IgE和IgG反应。

BACKGROUND & AIMS: BACKGROUND AND OBJECTIVES:Olaratumab is a recombinant human monoclonal antibody that binds to platelet-derived growth factor receptor-α (PDGFRα). In a randomized phase II study, olaratumab plus doxorubicin met its predefined primary endpoint for progression-free survival and achieved a highly significant improvement in overall survival versus doxorubicin alone in patients with advanced or metastatic soft tissue sarcoma (STS). In this study, we characterize the pharmacokinetics (PKs) of olaratumab in a cancer patient population. METHODS:Olaratumab was tested at 15 or 20 mg/kg in four phase II studies (in patients with nonsmall cell lung cancer, glioblastoma multiforme, STS, and gastrointestinal stromal tumors) as a single agent or in combination with chemotherapy. PK sampling was performed to measure olaratumab serum levels. PK data were analyzed by nonlinear mixed-effect modeling techniques using NONMEM®. RESULTS:The PKs of olaratumab were best described by a two-compartment PK model with linear clearance (CL). Patient body weight was found to have a significant effect on both CL and central volume of distribution (V 1), whereas tumor size significantly affected CL. A small subset of patients developed treatment-emergent anti-drug antibodies (TE-ADAs); however, TE-ADAs did not have any effect on CL or PK time course of olaratumab. There was no difference in the PKs of olaratumab between patients who received olaratumab as a single agent or in combination with chemotherapy. CONCLUSION:The PKs of olaratumab were best described by a model with linear disposition. Patient body weight and tumor size were found to be significant covariates. The PKs of olaratumab were not affected by immunogenicity or chemotherapeutic agents.

背景与目标：

BACKGROUND & AIMS: :M-type phospholipase A2 receptor (PLA2R) is the major target antigen in primary membranous nephropathy (PMN). Previous studies have evaluated the diagnostic value of serum anti-PLA2R antibody. However, the correlation of serum anti-PLA2R antibody and glomerular PLA2R deposition, and their association with clinical characteristics need to be further evaluated.A total of 136 patients were involved as inception group because serum anti-PLA2R antibody and glomerular PLA2R antigen were simultaneously measured. We examined serum anti-PLA2R antibody by ELISA and glomerular PLA2R deposition by immunofluorescence assay.Positive serum anti-PLA2R antibody and glomerular PLA2R deposition were seen in 58.8% (80/136) and 95.6% (130/136) patients, respectively (P < .001). Proteinuria, serum total protein, serum albumin, serum creatinine, and estimated glomerular filtration rate (eGFR) had significant differences between patients with serum anti-PLA2R antibody and those without. Serum anti-PLA2R antibody levels were correlated with serum albumin, serum creatinine, eGFR, and proteinuria. Glomerular PLA2R deposition intensities were weakly correlated with proteinuria. Unexpectedly, there was a positive correlation rather than a negative correlation between glomerular PLA2R deposition intensity and eGFR.In conclusion, serum anti-PLA2R antibody is more closely correlated with disease activity and renal function than glomerular PLA2R deposition.

背景与目标： : M型磷脂酶A2受体 (PLA2R) 是原发性膜性肾病 (PMN) 的主要靶抗原。先前的研究已经评估了血清anti-PLA2R抗体的诊断价值。但是，血清anti-PLA2R抗体与肾小球PLA2R沉积的相关性及其与临床特征的相关性有待进一步评估。共有136例患者作为起始组，因为同时测量了血清anti-PLA2R抗体和肾小球PLA2R抗原。ELISA法检测血清anti-PLA2R抗体，免疫荧光法检测肾小球PLA2R沉积，58.8% 例 (80/136例) 和95.6% 例 (130/136例) 血清anti-PLA2R抗体和肾小球PLA2R沉积阳性 (p  < .001)。蛋白尿，血清总蛋白，血清白蛋白，血清肌酐和估计的肾小球滤过率 (eGFR) 在具有血清anti-PLA2R抗体的患者与没有血清白蛋白的患者之间具有显着差异。血清anti-PLA2R抗体水平与血清白蛋白，血清肌酐，eGFR和蛋白尿相关。肾小球PLA2R沉积强度与蛋白尿的相关性较弱。不料，肾小球PLA2R沉积强度与eGFR呈正相关而非负相关，结论血清anti-PLA2R抗体与疾病活动和肾功能的相关性比肾小球PLA2R沉积更密切。

BACKGROUND & AIMS: :The host immunologic response to group A streptococcal infections gives rise to numerous antibodies directed against cellular and extracellular bacterial antigens. For determining individual immune status, or studying the pathogenesis of group A streptococcal associated diseases, such as acute rheumatic fever (ARF), an assay capable of determining antibodies responses to multiple antigens would be of great advantage. We have developed a microsphere based, multiplexed immunoassay for the simultaneous quantitation of antibodies to nine different extracellular, ARF related tissue and group A streptococci specific antigens using only 5 microl of sample. Through the selection of microspheres and serum diluent, non-specific antibody binding was reduced by 17%. Different formulations of the coupling buffer were found to greatly influence the efficiency of coupling antigens to the carboxylated microspheres. Monoclonal antibodies against the different antigens demonstrated assay specificity as well as sensitivities of less than 1 ng/ml of antibody. This multiplexed assay should be a powerful research and clinical tool in determining antibody responses to group A streptococcal infections and in potentially determining the role of a variety of cross-reactive antigens in rheumatic fever and rheumatic heart disease.

背景与目标： : 宿主对A组链球菌感染的免疫反应产生了许多针对细胞和细胞外细菌抗原的抗体。对于确定个体免疫状态或研究A组链球菌相关疾病 (例如急性风湿发热 (ARF)) 的发病机理，能够确定对多种抗原的抗体反应的测定法将具有很大的优势。我们已经开发了一种基于微球的多重免疫测定法，用于仅使用5 microl样品同时定量针对9种不同的细胞外，ARF相关组织和a组链球菌特异性抗原的抗体。经由过程选择微球和血清稀释剂，非特异性抗体结合被17% 削减。发现偶联缓冲液的不同配方会极大地影响将抗原偶联到羧化微球的效率。针对不同抗原的单克隆抗体显示出测定特异性以及小于1 ng/ml抗体的敏感性。这种多重分析应该是确定对a组链球菌感染的抗体反应以及潜在地确定多种交叉反应抗原在风湿发热和风湿性心脏病中的作用的强大研究和临床工具。

BACKGROUND & AIMS: We developed a direct microtiter plate enzyme immunoassay to measure estradiol-17 beta in saliva. The assay has a commercially available monoclonal antibody, raised against estradiol-17 beta-6-carboxymethyloximebovine serum albumin, and a homologous horseradish peroxidase conjugate measured colorimetrically. The detection limit (equivalent to B0-3 SD) is 365 amol/well or 7.3 pmol/L when 50-microL samples are assayed. Cross-reactivity with estrone and estriol, testosterone, or progesterone is < 0.2%. Estradiol-17 beta was measured in daily samples over five natural menstrual cycles and eight cycles stimulated as a preliminary to in vitro fertilization, and the concentrations and fluctuations found agreed with previously published data. This method gives results in approximately 3 h and may be useful for fertility monitoring and management.

背景与目标： 我们开发了一种直接微量滴定板酶免疫测定法来测量唾液中的estradiol-17 β。该测定法具有市售的抗estradiol-17 beta-6-carboxymethyloximebovine血清白蛋白的单克隆抗体和比色法测定的同源辣根过氧化物酶缀合物。当测定50μl样品时，检测限 (相当于B0-3 SD) 为365 amol/井或7.3 pmol/L。与雌酮和雌三醇、睾酮或孕酮的交叉反应性 <0.2%。Estradiol-17 β 在五个自然月经周期和八个周期刺激的每日样本中进行了体外受精的初步测定，发现的浓度和波动与先前发表的数据一致。此方法在大约3小时内得出结果，可能对生育监测和管理有用。

BACKGROUND & AIMS: BACKGROUND:The purpose of this study was to determine the distribution of radioiodinated estramustine (RI-EMP) and a radioiodinated antibody against estramustine binding protein (RI-EMBP-AB) in mice. METHODS:RI-EMP and RI-EMBP-AB were injected in male mice intravenously, and the activities of tissue samples were measured 1-31 hr from the injection. Pure iodine-125 (RI) was used as a control. RESULTS:RI-EMP accumulated in the prostate, which contained 2.6% of injected activity (ID) per gram tissue at 7 hr. The liver had an activity of 21.4% ID/g at 1 hr, which decreased as RI-EMP was secreted in bile. The lung contained 2.3% ID/g at 7 hr, and it retained the activity longer than the prostate. RI-EMBP-AB accumulated in the prostate: The activity was 2.9% ID/g at 7 hr. The gallbladder contained 6.5% ID/g at 7 hr. CONCLUSIONS:Due to its cytotoxic and radiosensitizing properties, RI-EMP can possibly be used for treating prostate cancer and other tumors.

背景与目标：

BACKGROUND & AIMS: :We have studied by electron microscopy a fascinating series of antidansyl monoclonal antibodies developed by Dangl et al. (Cytometry 2, 395-401, 1982) which have the same variable domain but different constant domains. Three of the four subclasses of mouse IgG were represented, IgG1, IgG2a and IgG2b. Previously, Oi et al. (Nature 307, 136-140, 1984) had examined the flexibilities of these antibodies by time-resolved fluorescence depolarization and found that IgG1 was least flexible, IgG2a was intermediate and IgG2b was the most flexible. In this communication we examine the conformations of circular complexes formed between these antibodies and a bivalent hapten, bis-dansyl cadaverine. The circular complexes were predominantly composed of two antibodies linked into a ring by two bivalent haptens, and are referred to as dimers, since only the antibody molecules are seen with the electron microscope. A few trimers and an occasional tetramer were also present in these preparations. For the least flexible IgG1, almost all (greater than 99%) of the circular dimers were "open-hinge" complexes with a hinge angle between the Fab arms of 100-120 degrees. For the intermediate IgG2a, most of the dimers were "open-hinge" complexes, but a larger percentage, 4 to 5%, had closed hinges with a hinge angle approaching 0 degrees. For the most flexible IgG2b, over 40% of the dimers were "closed-hinge" complexes. A model is proposed to explain these differences based upon orientation of the hapten in the combining site and differences in hinge structure.

背景与目标： : 我们通过电子显微镜研究了由Dangl等人开发的一系列令人着迷的抗dansyl单克隆抗体 (Cytometry 2，395-401，1982)，它们具有相同的可变结构域但具有不同的恒定结构域。代表了小鼠IgG的四个亚类中的三个，IgG1，IgG2a和IgG2b。以前，Oi等人 (Nature 307，136-140，1984) 通过时间分辨荧光去极化检查了这些抗体的柔韧性，发现IgG1是最不柔韧的，IgG2a是中间的，IgG2b是最柔韧的。在本交流中，我们检查了这些抗体与二价半抗原bis-dansyl尸胺之间形成的圆形复合物的构象。圆形复合物主要由两种通过两个二价半抗原连接成一个环的抗体组成，并且被称为二聚体，因为在电子显微镜下只能看到抗体分子。这些制剂中还存在一些三聚体和偶尔的四聚体。对于柔性最小的IgG1，几乎所有 (大于99%) 的圆形二聚体是 “开铰链” 复合物，其Fab臂之间的铰链角度为100-120度。对于中间体IgG2a，大多数二聚体是 “开铰链” 复合物，但较大的百分比 (4至5%) 具有铰链角度接近0度的闭合铰链。对于最灵活的IgG2b，超过40% 的二聚体是 “闭合铰链” 复合物。提出了一个模型来解释这些差异，该模型基于结合部位中半抗原的方向和铰链结构的差异。

BACKGROUND & AIMS: :Data are presented indicating that the growth of 5 out of 5 murine lymphoid tumors can be inhibited in a synergistic fashion in vitro by combined treatment with the iron chelator deferoxamine (DFO) and an immunoglobulin G (IgG) monoclonal anti-transferrin receptor antibody (ATRA). A two-way dose/response analysis shows that the ATRA becomes more efficient as an inhibitor with increasing doses of DFO. Flow cytometric studies further support the view that IgG ATRAS impair transferrin receptor (TR) function by causing TR down-modulation and degradation, even when the presence of DFO acts to promote increased cell surface TR expression. It is also shown that an IgG ATRA is nearly as effective as an IgM ATRA in inhibiting tumor cell growth when used in combination with DFO. Finally, studies with the iron chelator picolinic acid show that it produces only additive, or very slightly supra-additive, effects when used in combination with the ATRA. Therefore, these studies not only continue to suggest that combination chelator/ATRA therapy warrants further investigation as a tool in the therapy of hematopoietic malignancies, but also make the following new points: (1) the clinically familiar iron chelator deferoxamine, but not all iron chelators, produces synergistic inhibition of tumor growth in vitro with ATRAS; and (2) IgG ATRAS, which may be clinically more attractive reagents than IgA or IgM ATRAS because of better access to extra vascular tissue spaces, have unexpectedly been found to function as powerful growth inhibitors when used in combination with DFO.

背景与目标： : 提供的数据表明，通过与铁螯合剂去铁胺 (DFO) 和免疫球蛋白G (IgG) 单克隆抗转铁蛋白受体联合治疗，可以在体外以协同方式抑制5种鼠淋巴样肿瘤中的5种的生长抗体 (ATRA)。双向剂量/反应分析表明，随着DFO剂量的增加，ATRA作为抑制剂变得更有效。流式细胞术研究进一步支持以下观点: 即使DFO的存在促进细胞表面TR表达增加，IgG ATRAS也会通过引起TR下调和降解而损害运铁蛋白受体 (TR) 的功能。还显示，当与DFO组合使用时，IgG ATRA在抑制肿瘤细胞生长方面几乎与IgM ATRA一样有效。最后，对铁螯合剂吡啶甲酸的研究表明，与ATRA结合使用时，它仅产生添加剂或非常轻微的超添加剂效应。因此，这些研究不仅继续表明，螯合剂/ATRA联合治疗值得进一步研究作为治疗造血系统恶性肿瘤的工具，而且还提出了以下新的观点 :( 1) 临床上熟悉的铁螯合剂去铁胺，但不是所有的铁螯合剂，与ATRAS在体外产生肿瘤生长的协同抑制; 和 (2) IgG ATRAS在临床上可能比IgA或IgM ATRAS更具吸引力，因为更好地进入额外的血管组织空间，当与DFO组合使用时，已意外地发现IgG ATRAS具有强大的生长抑制剂的功能。

BACKGROUND & AIMS: :Oncolytic viruses exploit the cancer cell phenotype to complete their lytic life cycle, releasing progeny virus to infect nearby cells and repeat the process. We modified the oncolytic group B adenovirus EnAdenotucirev (EnAd) to express a bispecific single-chain antibody, secreted from infected tumour cells into the microenvironment. This bispecific T-cell engager (BiTE) binds to EpCAM on target cells and cross-links them to CD3 on T cells, leading to clustering and activation of both CD4 and CD8 T cells. BiTE transcription can be controlled by the virus major late promoter, limiting expression to cancer cells that are permissive for virus replication. This approach can potentiate the cytotoxicity of EnAd, and we demonstrate using primary pleural effusions and peritoneal malignant ascites that infection of cancer cells with the BiTE-expressing EnAd leads to activation of endogenous T cells to kill endogenous tumour cells despite the immunosuppressive environment. In this way, we have armed EnAd to combine both direct oncolysis and T cell-mediated killing, yielding a potent therapeutic that should be readily transferred into the clinic.

背景与目标： : 溶瘤病毒利用癌细胞表型来完成其裂解生命周期，释放后代病毒以感染附近的细胞并重复该过程。我们修改了溶瘤组B腺病毒EnAdenotucirev (EnAd) 表达双特异性单链抗体，该抗体从感染的肿瘤细胞分泌到微环境中。这种双特异性T细胞接合器 (BiTE) 与靶细胞上的EpCAM结合，并将它们与T细胞上的CD3交联，从而导致CD4和CD8 T细胞的聚集和激活。BiTE转录可以由病毒主要的晚期启动子控制，从而限制了对允许病毒复制的癌细胞的表达。这种方法可以增强EnAd的细胞毒性，并且我们证明使用原发性胸腔积液和腹膜恶性腹水，尽管存在免疫抑制环境，但用BiTE表达的EnAd感染癌细胞会导致内源性T细胞激活以杀死内源性肿瘤细胞。通过这种方式，我们已经武装EnAd将直接溶解和T细胞介导的杀伤结合起来，产生了一种有效的治疗方法，可以很容易地转移到临床中。

BACKGROUND & AIMS: :Successful translation of findings derived from preclinical studies into effective therapies is critical in biomedical research. Lack of robustness and reproducibility of the preclinical data, due to insufficient number of repeats, inadequate cell-based and mouse models contribute to the poor success rate. Antibodies are widely used in preclinical research, notably to determine the expression of potential therapeutic targets in tissues of interest, including tumors, but also to identify disease and/or treatment response biomarkers. We sought to determine whether the current antibody characterization standards in preclinical research are sufficient to ensure reliability of the data found in peer-reviewed publications. To address this issue, we used detection of the protein c-FLIP, a major factor of resistance to apoptosis, as a proof of concept. Accurate detection of endogenous c-FLIP levels in the preclinical settings is imperative since it is considered as a potential theranostic biomarker. Several sources of c-FLIP antibodies validated by their manufacturer and recommended for western blotting were therefore rigorously tested. We found a wide divergence in immune recognition properties. While these antibodies have been used in many publications, our results show that several of them failed to detect endogenous c-FLIP protein by Western blotting. Our results suggest that antibody validation standards are inadequate, and that systematic use of genetic knockdowns and/or knockouts to establish proof of specificity is critical, even for antibodies previously used in the scientific literature. Because antibodies are fundamental tools in both preclinical and clinical research, ensuring their specificity is crucial.

背景与目标： : 成功地将临床前研究的发现转化为有效的疗法在生物医学研究中至关重要。由于重复次数不足，基于细胞的和小鼠模型不足，缺乏临床前数据的稳健性和可重复性导致成功率低。抗体广泛用于临床前研究，特别是用于确定感兴趣组织 (包括肿瘤) 中潜在治疗靶标的表达，还用于鉴定疾病和/或治疗反应生物标志物。我们试图确定临床前研究中当前的抗体表征标准是否足以确保同行评审出版物中发现的数据的可靠性。为了解决这个问题，我们使用了检测蛋白c-FLIP (抗凋亡的主要因素) 作为概念的证明。临床前环境中内源性c-FLIP水平的准确检测势在必行，因为它被认为是潜在的肿瘤生物标志物。因此，对其制造商验证并推荐用于蛋白质印迹的c-FLIP抗体的几种来源进行了严格测试。我们发现免疫识别特性存在很大差异。尽管这些抗体已在许多出版物中使用，但我们的结果表明，其中一些未能通过Western印迹检测到内源性c-FLIP蛋白。我们的结果表明，抗体验证标准是不充分的，并且系统地使用基因敲除和/或敲除来建立特异性证明是至关重要的，即使对于以前在科学文献中使用的抗体也是如此。由于抗体是临床前和临床研究的基本工具，因此确保其特异性至关重要。

BACKGROUND & AIMS: :Although many multiple myeloma (MM) patients initially respond to cytotoxic therapy, most eventually relapse. Novel therapeutic strategies employing a combination of chemotherapy with targeted biologics may significantly enhance the response of tumor cells to treatment. We tested a fully human anti-IGF-IR antibody (A12) against MM, and showed specific inhibition of IGF-I or serum-induced IGF-IR signaling in MM cells in vitro. The A12 as a single agent was demonstrated to exert modest to significant inhibition of tumor growth in vivo in various subcutaneous xenograft MM models. The A12 was also evaluated in a disseminated xenograft MM.1S NOD/SCID model as monotherapy or in combination with other drugs (bortezomib, melphalan) currently in clinical use. The tumor burden, as determined by luciferase bioimaging, was sharply decreased, and overall survival significantly prolonged when the therapies were combined. Immunohistochemical analysis demonstrated that the A12 treated tumors had significantly decreased vascularization compared to control tumors. Furthermore, most MM lines constitutively secreted significant quantities of VEGF, and this was enhanced following IGF-I treatment. Inhibition of IGF-IR by the A12 in vitro suppressed both constitutive and IGF-I-induced secretion of VEGF, indicating that a putative anti-angiogenic mechanism associated with the A12 treatment may contribute to its anti-tumor effect.

背景与目标： : 尽管许多多发性骨髓瘤 (MM) 患者最初对细胞毒性治疗有反应，但大多数最终复发。采用化疗与靶向生物制剂相结合的新型治疗策略可能会显着增强肿瘤细胞对治疗的反应。我们测试了针对MM的全人抗igf-ir抗体 (A12)，并在体外显示了对MM细胞中igf-i或血清诱导的igf-ir信号的特异性抑制。在各种皮下异种移植MM模型中，A12作为单一药物被证明对体内肿瘤生长具有适度至显着的抑制作用。还在播散性异种移植MM.1S NOD/SCID模型中评估了A12，作为单一疗法或与目前临床使用的其他药物 (硼替佐米，美法仑) 联合使用。通过荧光素酶生物成像确定的肿瘤负荷急剧降低，并且当联合治疗时，总生存期显着延长。免疫组织化学分析表明，与对照肿瘤相比，A12治疗的肿瘤血管形成明显减少。此外，大多数MM系组成型分泌了大量的VEGF，并且在igf-i治疗后这种情况得到了增强。体外A12抑制igf-ir抑制了组成型和igf-i诱导的VEGF分泌，表明与A12治疗相关的推定抗血管生成机制可能有助于其抗肿瘤作用。

BACKGROUND & AIMS: :A major traditional of antibacterial drugs is antibiotic which promotes more rapid release of the toxins from bacteria cells in human body, which causes severe infection. The thermostable direct hemolysin (TDH) has been proposed as a major virulence factor of Vibrio parahaemolyticus (Vp). This study covers the preparation of polymer microparticle-antibody conjugate for the development of a drug targeting approach for antibacterial drug delivery. The chemical binding of antibodies (ab) to latex bead of 0.2 mum diameter was performed by using a water-soluble carbodiimide technique. Confocal microscopy revealed that the bacteria were strongly absorbed by the latex beads with bound anti-Vp polyclonal antibody (pAb). Treatment with a latex bead bound both anti-Vp pAb and anti-TDH monoclonal antibody (mAb) significantly inhibited bacterial adherence to the Caco-2 cells (p < 0.01), and reduced TDH-induced cytotoxicity in histology. These preliminary results suggest that it may be possible to effectively protect against Vp infection by using this microparticle-antibody conjugate delivery system.

背景与目标： 抗菌药物的主要传统是抗生素，它促进人体细菌细胞中毒素的更快释放，从而引起严重的感染。已提出热稳定的直接溶血素 (TDH) 作为副溶血性弧菌 (Vp) 的主要毒力因子。这项研究涵盖了聚合物微粒-抗体偶联物的制备，用于开发用于抗菌药物递送的药物靶向方法。通过使用水溶性碳二亚胺技术进行抗体 (ab) 与直径为0.2的乳胶珠的化学结合。共聚焦显微镜显示，细菌被结合了抗Vp多克隆抗体 (pAb) 的乳胶珠强烈吸收。用结合抗Vp pAb和抗TDH单克隆抗体 (mAb) 的乳胶珠处理可显着抑制细菌对Caco-2细胞的粘附 (p <0.01)，并降低组织学中TDH诱导的细胞毒性。这些初步结果表明，通过使用这种微粒-抗体缀合物递送系统，有可能有效地预防Vp感染。

BACKGROUND & AIMS: :Intravenous injections into nude mice of 5 mg/kg methotrexate (MTX) linked to the antibody to human high molecular weight-melanoma associated antigen (HMW-MAA), monoclonal antibody (mAb) 225.28, an IgG2a, on days 1, 4, 7, 10 and 14, starting 24 h after subcutaneous inoculation of 2 x 10(6) cultured human M21 melanoma cells inhibited mean tumor volume by 90% on day 14 and by 65% on day 50 after the beginning of the treatment. Injections of equimolar amounts of free MTX and MTX linked to normal mouse IgG or to an isotype-matched myeloma protein did not inhibit tumor growth significantly. MTX linked to mAb 225.28 did not inhibit the xenograft of a subline of human melanoma cell line M21 without detectable expression of HMW-MAA. In a clonogenic assay, the MTX-225.28 conjugate was three times more potent in inhibiting the growth of M21 melanoma cells than free MTX, but did not inhibit the growth of kidney carcinoma cells Caki-1, which do not express high-Mr MAA. In contrast, MTX linked to the mAb DAL K29, reacting with kidney carcinoma cells Caki-1, inhibited their growth but did not affect that of melanoma cells. M21 melanoma cells isolated from the residual tumor of a mouse treated with the MTX-225.28 conjugate did not differ in their reactivity with mAb 225.28 and in their sensitivity to MTX when compared with M21 cells from an untreated mouse.

背景与目标： : 在第1、4、7、10和14天，将5 mg/kg甲氨蝶呤 (MTX) 与人高分子量黑色素瘤相关抗原 (hmw-maa)，单克隆抗体 (mAb) 225.28 (IgG2a) 连接的抗体静脉注射到裸鼠中，在皮下接种2 × 10(6) 培养的人M21黑色素瘤细胞后24小时开始，在治疗开始后第14天90% 抑制平均肿瘤体积，并在第50天65% 抑制平均肿瘤体积。注射等摩尔量的游离MTX和MTX与正常小鼠IgG或同种型匹配的骨髓瘤蛋白连接不会显着抑制肿瘤生长。与mAb 225.28连接的MTX不抑制人黑素瘤细胞系M21的亚系的异种移植，而没有可检测到的hmw-maa表达。在克隆形成测定中，MTX-225.28缀合物在抑制M21黑色素瘤细胞生长方面的效力是游离MTX的三倍，但Caki-1不抑制不表达高Mr MAA的肾癌细胞的生长。相反，与mAb DAL K29连接的MTX与Caki-1的肾癌细胞反应，抑制了它们的生长，但不影响黑色素瘤细胞的生长。与未处理小鼠的M21细胞相比，从用MTX-225.28缀合物处理的小鼠的残留肿瘤中分离的M21黑色素瘤细胞在与mAb 225.28的反应性和对MTX的敏感性方面没有差异。

BACKGROUND & AIMS: :Internalization by cells of radiolabeled protein ligands bound to the cell surface is frequently analyzed by extraction of the cells with low pH buffers. This treatment supposedly strips the ligands from the cell surface, and remaining molecules are considered to be internalized. However, we show herein that: (1) low molecular weight catabolic products that are trapped within lysosomes (residualizing radiolabels) are efficiently extracted by low pH buffers, under the same conditions used to remove cell surface-bound material, and (2) low pH treatment lyses the majority of the cells, as shown with both a nonadherent and an adherent cell line, with the release of most of a (51)Cr label. Still, low pH extraction was effective at demonstrating Ab internalization, as has been demonstrated many times. These effects of low pH treatment may be attributed to the fixative properties of these buffers. Regardless of the mechanism, these data must be taken into consideration in interpreting the results of such experiments.

背景与目标： : 通过细胞内化结合到细胞表面的放射性标记蛋白配体经常通过用低pH缓冲液提取细胞来分析。据推测，这种处理将配体从细胞表面剥离，剩余的分子被认为是内在化的。然而，我们在本文中显示 :( 1) 在用于去除细胞表面结合材料的相同条件下，通过低pH缓冲液有效地提取了溶酶体中捕获的低分子量分解代谢产物 (残留放射性标记)，以及 (2) 低pH处理裂解大多数细胞，如非粘附细胞系和粘附细胞系所示，大部分 (51)Cr标记的释放。尽管如此，低pH萃取仍可有效证明Ab内化，正如多次证明的那样。低pH处理的这些影响可能归因于这些缓冲液的固定性。无论机制如何，在解释此类实验结果时必须考虑这些数据。